High-speed countercurrent chromatography in plants
Extracting and purifying active ingredients from plants is a necessary prerequisite for rational formulation, efficacy research, and structural identification. However, the content of active ingredients in most plants is low, making it difficult to enrich; the system is complex, with macromolecules and small molecules, living and non-living substances coexisting, especially the existence of isomers with similar structures, making separation and purification difficult. With the advancement of the modernization and standardization process of traditional Chinese medicine,chemical referenceThe traditional standard reference substance preparation method is time-consuming, laborious, low yield, low purity, high cost and complicated process [1]. Therefore, it is necessary to strengthen the basic research of natural product extraction and separation, develop new efficient, inexpensive, simple and high-quality separation and purification technologies, and prepare more natural product chemical reference substances to meet the needs of traditional Chinese medicine modernization research.
High-speed countercurrent chromatography (HSCCC) is a carrier-free liquid-liquid distribution chromatography. Its separation principle is to use the unidirectional hydrodynamic equilibrium principle to make two mutually insoluble solvent phases unidirectionally distributed in a high-speed rotating spiral tube. One of the phases is used as a stationary phase, and the mobile phase carrying the sample is transported through the stationary phase by a constant current pump, and the separation is realized by the difference of the partitioning coefficient (K) of the sample in the two phases. There are many advantages that cannot be compared with traditional liquid-solid chromatography. It completely excludes the irreversible adsorption, staining, denaturation and inactivation of the support on the sample. In addition, its loading amount is relatively large, up to a few grams, which is 104 to 105 times that of high-performance liquid chromatography; compared with atmospheric and low-pressure chromatography, although the loading amount is small, the separation ability is much better. Some samples can obtain one or even more monomers after one separation, and the separation time is short, usually a few hours to complete a separation [2, 3]. It can be seen that countercurrent chromatography technology is inNatural plant extractsThe preparation of standard products has unique advantages.
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